Esculetin inhibits cartilage resorption induced by interleukin 1alpha in combination with oncostatin M
| Autor: | SF Elliott, S. Carrère, Andrew D. Rowan, P. J. T. Koshy, Tim E. Cawston, JB Catterall |
|---|---|
| Rok vydání: | 2001 |
| Předmět: |
Immunology
Enzyme-Linked Immunosorbent Assay Matrix metalloproteinase General Biochemistry Genetics and Molecular Biology Hydroxyproline chemistry.chemical_compound Chondrocytes Rheumatology medicine Immunology and Allergy Animals Humans Umbelliferones Cells Cultured Biological Products Tissue Inhibitor of Metalloproteinase-1 biology Dose-Response Relationship Drug Cartilage Oncostatin M Blotting Northern Molecular biology Matrix Metalloproteinases Resorption Methylene Blue Extended Report medicine.anatomical_structure Proteoglycan chemistry Cell culture biology.protein Collagenase Cattle Proteoglycans Collagen medicine.drug Interleukin-1 |
| Zdroj: | Annals of the rheumatic diseases. 60(2) |
| ISSN: | 0003-4967 |
| Popis: | OBJECTIVE—To determine if a new inhibitor, esculetin (EST), can block resorption of cartilage. METHODS—Interleukin 1α (IL1α, 0.04-5 ng/ml) and oncostatin M (OSM, 0.4-50 ng/ml) were used to stimulate the release of proteoglycan and collagen from bovine nasal cartilage and human articular cartilage in explant culture. Proteoglycan and collagen loss were assessed by dimethylmethylene blue and hydroxyproline assays, respectively. Collagenase levels were measured by assay of bioactivity and by enzyme linked immunosorbent assay (ELISA). The effects of EST on the expression of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in the transformed human chondrocyte cell line T/C28a4 were assessed by northern blot analysis. TIMP-1 protein levels were assayed by ELISA. The effect of EST on the MMP-1 promoter was assessed using a promoter-luciferase construct in transient transfection studies. RESULTS—EST inhibited proteoglycan and collagen resorption in a dose dependent manner with significant decreases seen at 66 µM and 100 µM EST, respectively. Collagenolytic activity was significantly decreased in bovine nasal cartilage cultures. In human articular cartilage, EST also inhibited IL1α + OSM stimulated resorption and decreased MMP-1 levels. TIMP-1 levels were not altered compared with controls. In T/C28a4 chondrocytes the IL1α + OSM induced expression of MMP-1, MMP-3, and MMP-13 mRNA was reduced to control levels by 250 µM EST. TIMP-1 mRNA levels were unaffected by EST treatment. All cytokine stimulation of an MMP-1 luciferase-promoter construct was lost in the presence of the inhibitor. CONCLUSION—EST inhibits degradation of bovine nasal cartilage and human articular cartilage stimulated to resorb with IL1α + OSM. |
| Databáze: | OpenAIRE |
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